Isolation and identification of the cyanotoxin cylindrospermopsin and deoxy-cylindrospermopsin from a Thailand strain of Cylindrospermopsis raciborskii (Cyanobacteria)

A strain of Cylindrospermopsis (Cyanobacteria) isolated from a fishpond in Thailand was examined for its taxonomy based upon morphology and 16S rRNA g ene sequence. It was also examined for production of the hepatotoxic cyanot oxin called cylindrospermopsin (CYN) and deoxycylindrospermopsin (deoxy-CYN ). The strain (CY-Thai) was identified as C. raciborskii (Woloszynska) Seen aya and Subba Raju based upon morphological examination which was confirmed by 16S rRNA gene sequences and phylogenetic comparisons based upon its 16S rRNA gene. The alkaloid heptatotoxin CYN was confirmed using mouse bioassa y, HPLC and HPLC-MS/MS while deoxy-CYN was confirmed using HPLC-MS/MS. The mouse bioassay gave a minimum lethal dose at 250 mg dry weight cells/kg bod y weight within 24 h and 125 mg/kg at 72 h, with signs of poisoning the sam e as in literature reports for CYN. HPLC chromatographic comparison of the CY-Thai toxin with standard CYN gave the same retention time and an absorba nce maximum at 262 nm. HPLC-MS/MS confirmed the presence of CYN (M + H 416) and deoxy-CYN (M + H 400). The CYN content in strain CY-Thai was estimated at 1.02 mg/g and approximately 1/10 of this amount for deoxy-CYN. This is the first report from Asia of a CYN, deoxy-CYN producing Cylindrospermopsis raciborskii. (C) 2001 Elsevier Science Ltd. All rights reserved.